Novel Smallpox Vaccine Derived from VV/VAR Immunome

描述(由申请人提供)：EpiVax通过筛选候选疫苗成分的全基因组，专门开发表位驱动的疫苗。在这项应用中，我们建议基于牛痘病毒(VV)和天花(VAR)免疫小体之间保存的表位开发一种安全的新天花疫苗，在发生生物恐怖袭击时可用作预防和治疗干预。首先，将通过计算机驱动(EpiMatrix)分析来筛选在痘苗病毒(W)和天花(Var)基因组之间保守的多肽序列以寻找免疫优势表位，并使用来自VV免疫个体的T细胞进行确认。其次，选定的表位将被比对并克隆到优化的DNA疫苗载体中，以便在体内高效和持续表达。第三，我们将用DNA疫苗构建物接种人类白细胞抗原转基因小鼠，使用细胞内细胞因子流式细胞术测量对DNA疫苗构建体的从头免疫反应。EpiVax将负责选择Vv/Var表位并管理该项目。布朗大学的结核病/艾滋病毒研究实验室(分包商)将进行结合分析，并开发由选定表位串组成的寡核苷酸多表位结构。克罗博士的实验室(分包商，范德比尔特大学)将进行ELISpot分析。韦纳博士(宾夕法尼亚大学Stella Chance实验室分包商)将进行疫苗接种研究。第一阶段的最终产品将是对几种原型天花疫苗构建物在人类白细胞抗原转基因小鼠中的免疫原性和保护效果的评估。在第二阶段，我们将筛选更多的表位，构建最终的疫苗构建物，通过佐剂和输送工具优化最终疫苗构建物的免疫原性，并解决这些构建物在人体中的安全性、毒性和免疫原性。

DESCRIPTION (provided by applicant): EpiVax specializes in the development of epitope-driven vaccines by screening whole genomes for candidate vaccine components. In this application, we propose to develop a safe, new smallpox vaccine based on epitopes conserved between the vaccinia virus (VV) and Variola (VAR) 'immunomes' that could be used as both a prophylactic and a therapeutic intervention in the event of a bioterrorist attack. First, peptide sequences that are conserved between the vaccinia virus (W) and Variola (Var) genomes will be screened for immunodominant epitopes by computer-driven (EpiMatrix) analysis and confirmed using T cells from VV-immunized individuals. Second, selected epitopes will then be aligned and cloned into DNA vaccine vectors optimized for strong and sustained expression in vivo. Third, we will vaccinate HLA-transgenic mice with the DNA vaccine constructs, measuring de novo immune responses to the DNA vaccine constructs using intracellular cytokine flow cytometry. EpiVax will be responsible for selecting the VV/Var epitopes and managing the project. The TB/HIV Research Lab at Brown University (subcontractor) will perform binding assays and develop the oligonucleotide multi-epitope constructs consisting of strings of the selected epitopes. Dr. Crowe's laboratory (subcontractor, Vanderbilt University) will perform ELISpot assays. Dr. Weiner (subcontractor, Stellar Chance Laboratories, University of Pennsylvania) will perform the vaccination studies. The final product of Phase I will be an evaluation of the immunogenicity and protective effect of several prototype smallpox vaccine constructs in HLA transgenic mice. In Phase II we would screen additional epitopes, build the final vaccine constructs, optimize the immunogenicity of the final vaccine constructs with adjuvants and delivery vehicles, and address the safety, toxicity and immunogenicity of the constructs in human subjects.